Glutathione-protein mixed disulfide decreases the affinity of rat liver fatty acid-binding protein for unsaturated fatty acid.
.16 +/- 0.062% of the fatty acid-binding protein purified from 50 mM N-ethylmaleimide-treated rat liver (L-FABP) was determined as a form S-thiolated by glutathione (L-FABP-SSG). L-FABP-SSG, which was prepared in vitro through thiol-disulfide exchange reaction, showed more acidic pI (approximately 5.0) than the pI (approximately 7.0) of reduced L-FABP. S-thiolation of L-FABP by glutathione decreased the affinity of the protein for unsaturated fatty acids without changing the equimolar maximum binding. The changes in Kd were from 0.63 +/- 0.054 microM to 1.03 +/- 0.14 microM for oleic acid, from 0.63 +/- 0.028 microM to 0.97 +/- 0.12 microM for linoleic acid and from 0.85 +/- 0.050 microM to 1.45 +/- 0.024 microM for arachidonic acid. This modification did not alter the affinity nor the maximum binding for saturated fatty acids, which were determined to be Kd of approximately 1.0 microM for palmitic acid and approximately 0.9 microM for stearic acids, and equimolar maximum binding for both fatty acids. The binding affinity of L-FABP for unsaturated fatty acid may be regulated by redox state of the liver.
Hitomi
M
M
Department of Biochemistry, Niigata University School of Medicine, Japan.
Odani
S
S
Ono
T
T
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001665:Binding Sites / Q000187:drug effects; D002352:Carrier Proteins / Q000032:analysis / Q000378:metabolism*; D002850:Chromatography, Gel; D004220:Disulfides / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D005033:Ethylmaleimide / Q000494:pharmacology; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D005231:Fatty Acids, Unsaturated / Q000032:analysis / Q000378:metabolism*; D005978:Glutathione / Q000494:pharmacology*; D007700:Kinetics; D008099:Liver / Q000378:metabolism*; D008297:Male; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D051381:Rats; D011919:Rats, Inbred Strains; D013438:Sulfhydryl Compounds / Q000032:analysis
D000596:Amino Acids; D002352:Carrier Proteins; D004220:Disulfides; C493320:Fabp1 protein, mouse; C493321:Fabp1 protein, rat; C490174:Fabp7 protein, rat; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D005231:Fatty Acids, Unsaturated; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D013438:Sulfhydryl Compounds; D005978:Glutathione; D005033:Ethylmaleimide